Evidence that a burst of DNA depurination in SENCAR mouse skin induces error-prone repair and forms mutations in the H-ras gene

Dhrubajyoti Chakravarti, Paula C. Mailander, Kai Ming Li, Sheila Higginbotham, Henry L. Zhang, Michael L. Gross, Jane L Meza, Ercole Cavalieri, Eleanor G Rogan

Research output: Contribution to journalArticle

167 Citations (Scopus)

Abstract

Treatment of SENCAR mouse skin with dibenzo[a, l]pyrene results in abundant formation of abasic sites that undergo error-prone excision repair, forming oncogenic H-ras mutations in the early preneoplastic period. To examine whether the abundance of abasic sites causes repair infidelity, we treated SENCAR mouse skin with estradiol-3, 4-quinone (E 2 -3, 4-Q) and determined adduct leveis 1 h after treatment, as well as mutation spectra in the H-ras gene between 6 h and 3 days after treatment. E 2 -3, 4-Q formed predominantly (≥99%) the rapidly-depurinating 4-hydroxy estradiol (4-OHE 2 )-1-N3Ade adduct and the slower-depurinating 4-OHE 2 -1-N7Gua adduct. Between 6 h and 3 days, E 2 -3, 4-Q induced abundant A to G mutations in H-ras DNA, frequently in the context of a 3′-G residue. Using a T.G-DNA glycosylase (TDG)-PCR assay, we determined that the early A to G mutations (6 and 12 h) were in the form of G.T heteroduplexes, suggesting misrepair at A-specific depurination sites. Since G-specific mutations were infrequent in the spectra, it appears that the slow rate of depurination of the N7Gua adducts during active repair may not generate a threshold level of G-specific abasic sites to affect repair fidelity. These results also suggest that E 2 -3, 4-Q, a suspected endogenous carcinogen, is a genotoxic compound and could cause mutations.

Original languageEnglish (US)
Pages (from-to)7945-7953
Number of pages9
JournalOncogene
Volume20
Issue number55
DOIs
StatePublished - Nov 29 2001

Fingerprint

Inbred SENCAR Mouse
ras Genes
Skin
Mutation
DNA
DNA Glycosylases
DNA Repair
Carcinogens
Estradiol
Polymerase Chain Reaction

Keywords

  • Abasic sites
  • DNA repair
  • Depurinating adducts
  • Estradiol-3, 4-quinone
  • H-ras mutation in SENCAR mouse skin
  • Mismatched heteroduplex

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

Evidence that a burst of DNA depurination in SENCAR mouse skin induces error-prone repair and forms mutations in the H-ras gene. / Chakravarti, Dhrubajyoti; Mailander, Paula C.; Li, Kai Ming; Higginbotham, Sheila; Zhang, Henry L.; Gross, Michael L.; Meza, Jane L; Cavalieri, Ercole; Rogan, Eleanor G.

In: Oncogene, Vol. 20, No. 55, 29.11.2001, p. 7945-7953.

Research output: Contribution to journalArticle

Chakravarti, Dhrubajyoti ; Mailander, Paula C. ; Li, Kai Ming ; Higginbotham, Sheila ; Zhang, Henry L. ; Gross, Michael L. ; Meza, Jane L ; Cavalieri, Ercole ; Rogan, Eleanor G. / Evidence that a burst of DNA depurination in SENCAR mouse skin induces error-prone repair and forms mutations in the H-ras gene. In: Oncogene. 2001 ; Vol. 20, No. 55. pp. 7945-7953.
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abstract = "Treatment of SENCAR mouse skin with dibenzo[a, l]pyrene results in abundant formation of abasic sites that undergo error-prone excision repair, forming oncogenic H-ras mutations in the early preneoplastic period. To examine whether the abundance of abasic sites causes repair infidelity, we treated SENCAR mouse skin with estradiol-3, 4-quinone (E 2 -3, 4-Q) and determined adduct leveis 1 h after treatment, as well as mutation spectra in the H-ras gene between 6 h and 3 days after treatment. E 2 -3, 4-Q formed predominantly (≥99{\%}) the rapidly-depurinating 4-hydroxy estradiol (4-OHE 2 )-1-N3Ade adduct and the slower-depurinating 4-OHE 2 -1-N7Gua adduct. Between 6 h and 3 days, E 2 -3, 4-Q induced abundant A to G mutations in H-ras DNA, frequently in the context of a 3′-G residue. Using a T.G-DNA glycosylase (TDG)-PCR assay, we determined that the early A to G mutations (6 and 12 h) were in the form of G.T heteroduplexes, suggesting misrepair at A-specific depurination sites. Since G-specific mutations were infrequent in the spectra, it appears that the slow rate of depurination of the N7Gua adducts during active repair may not generate a threshold level of G-specific abasic sites to affect repair fidelity. These results also suggest that E 2 -3, 4-Q, a suspected endogenous carcinogen, is a genotoxic compound and could cause mutations.",
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AU - Chakravarti, Dhrubajyoti

AU - Mailander, Paula C.

AU - Li, Kai Ming

AU - Higginbotham, Sheila

AU - Zhang, Henry L.

AU - Gross, Michael L.

AU - Meza, Jane L

AU - Cavalieri, Ercole

AU - Rogan, Eleanor G

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